Pituitary metastasis from renal cell carcinoma: case report and review of the literature.

Pituitary metastasis(PM) from renal cell carcinoma(RCC) is rare, and is easy to be misdiagnosed. Here, we present a case of pituitary metastasis from clear-cell renal cell carcinoma(ccRCC) which was difficult to distinguish from other sellar region tumors. In addition, we systematically review the literature to find the characteristics of different tumors of the sellar region. It provides a new idea for the diagnosis of sellar region tumors in the clinic.

A good choice for the patients with prior failed ventriculoperitoneal shunt treatment of suprasellar arachnoid cysts: endoscopic fenestration.

To investigate the effectiveness of endoscopic fenestration in the patients with prior failed ventriculoperitoneal (VP) shunt treatment of suprasellar arachnoid cysts (SACs). Between 2012 and 2018, four pediatric patients of SACs with previous failed VP shunt treatment were surgically treated using endoscopic ventriculocystocisternostomy (VCC) in our hospital. The clinical symptoms, imaging data, and surgical outcomes were collected and analyzed retrospectively. A literature review is provided with regard to the reasons of shunt failure and surgical outcome of further endoscopic fenestration in the previously reported patients of SACs with prior failed VP shunt. For the 4 cases, the initial clinical symptoms relieved or even disappeared after shunt placement, but, respectively, recurred 2, 6, 11, and 6 months later. MR scans were conducted when the clinical symptoms reappeared and showed a cyst had greatly enlarged after shunt placement. Furthermore, VP shunt-related slit ventricle was also demonstrated in 3 cases. Clinical improvement and cysts shrinkage occurred in all 4 patients after VCC. Slit ventricle and hydrocephalus were also resolved. Three patients had their shunt apparatus removed after VCC, and another patient's guardian refused to remove the shunt apparatus. Subdural hematoma occurred in one case after shunt apparatus removal. Four patients have been stable during follow-up period (mean follow-up 26.5 months). All the three patients whose VP shunt were removed were shunt independence. There were 24 patients who underwent endoscopic fenestration as an alternative to the failed VP shunt treatment in the published reports. Added our 4 patients to the published group, the effective rate of endoscopic fenestration for SACs following previous failed VP shunt treatment was approximately 93% (26/28). Of the 24 patients, the shunt apparatuses were in situ or reimplantation in 9 patients due to shunt dependence. The correction to recognize the SAC is the first condition to select the optimal management philosophy. The analysis of the series suggests endoscopic operation is still an effective and safe option in the SAC patients with previous failed VP shunt, and the shunt apparatus can be removed for some patients. The short interval time between shunt operation and endoscopic fenestration is conductive to return patients to the shunt-free state.

Integrative proteomics and transcriptomics identify novel invasive-related biomarkers of non-functioning pituitary adenomas.

Non-functioning pituitary adenomas (NFPAs) are usually macroadenomas and display invasion into surrounding tissues. The treatment for invasive NFPAs is still challenging. This study describes the differential patterns of gene expression between invasive and non-invasive NFPAs and identifies novel biomarkers involved in invasion of NFPAs for diagnosis and treatment. Using gene microarray technology, we examined the gene expression profile and found 1160 differentially expressed messenger RNA (mRNA) between invasive and non-invasive NFPAs. Then, we examined the protein profile by liquid chromatography tandem mass spectrometry (LC-MS/MS) and found 433 differentially expressed proteins between invasive and non-invasive NFPAs. Subsequently, we integrated the proteomics and transcriptomics datasets and identified 29 common changed molecules. Through bioinformatics analysis using Ingenuity Pathway Analysis (IPA) software, we showed that the 29 molecules were enriched in 25 canonical signaling pathways, 25 molecular and cellular functions, and 2 networks. Eight genes were identified involved in the invasion function by the molecular and cellular functions analysis, including CAT, CLU, CHGA, EZR, KRT8, LIMA1, SH3GLB2 and SLC2A1. Furthermore, we validated the decreased CHGA expression and increased CLU expression in invasive NFPAs by qRT-PCR and Western blot. Our study demonstrated that integration of proteomics and transcriptomics could prove advantageous for accelerating tumor biomarker discovery and CHGA and CLU might be important novel biomarkers and therapeutic targets for invasion of NFPAs.

[Association between differentiation potential of immortalized human mesenchymal stem cells and cluster of differentiation antigens].

OBJECTIVE: To explore the relationship between multi-differentiation potential of monoclonal immortalized human mesenchymal stem cells (hMSC-TERT) in vivo and their cluster of differentiation antigens (CD). METHODS: Monoclonal hMSC-TERT were isolated using limiting dilution. Direct immunofluorescence staining flow cytometry was used to detect the cluster of differentiation antigens (CD44, CD45, CD105) of these cell lines. Their adipocytic, osteogenic, neuronal differentiation potential in vitro were determined by Oil Red O staining, Von Kossa staining and immunocytochemistry for tubulin-ß III antibody. Then hMSC-TERTs were transplanted subcutaneously into severe combined immunodeficiency (SCID) mice. The cell grafts were removed and analyzed by immunohistochemistry for pathologic tissue markers for multi-differentiation potential of hMSC-TERT cells in vivo. RESULTS: CD105+ cell was 84.68% positive in hMSC-TERT-C19 while <5% in other monoclonal cell lines. The positive rate of cytokeratin in grafts formed by hMSC-TERT-C19 cell line in SCID mice was much higher than the others (P < 0.01). CONCLUSION: The positive CD105 of monoclonal hMSC-TERTs may be directly correlated with its epithelial differentiation potential.

[Effects of high plating densities on in vitro and in vivo differentiation capability of immortalized human mesenchymal stem cells].

OBJECTIVE: To determine whether different cell plating densities could influence the potential differentiation of immortalized human mesenchymal stem cells (hMSC-TERT). METHODS: Extensive characterization was performed for two independent cell lines derived from parental hMSC-TERT cell line based on different plating densities during expansion in culture: 1: 2 (hMSC-TERT2) and 1: 20 (hMSC-TERT20). Their adipocytic, osteogenic, neuronal differentiation potential in vitro and multidirectional differentiation potential in vivo were analyzed by immunohistochemistry for pathologic tissue markers. RESULTS: hMSC-TERT2 formed cellular adipose foci and mineralized tuberculum after in vitro induction while hMSC-TERT20 showed adipocytes but no adipose foci or mineralized tuberculum. The positive rate of hMSC-TERT2 formed tissues on VIM, GFAP and LCA (89.17% ± 5.97%, 72.5% ± 10.11% and 76.67% ± 11.15%) under SCID murine skin were more than that of hMSC-TERT20 formed tissues (0.33% ± 0.65%, 9.58% ± 4.29% and 22.08% ± 6.95%). CONCLUSION: The cell lines derived at high plating density may have more powerful differentiation potential in vitro and in vivo than those derived at low cell plating density.

Endoscopic Fenestration for Treating Galassi Type III Middle Cranial Fossa Arachnoid Cysts: Single- and Multiple-stoma have the Same Curative Effect.

BACKGROUND: For endoscopic fenestration of middle cranial fossa arachnoid cysts (MCFACs), the decisions on the location and number of stomas are key issues. However, research on this particular topic has been limited. Thus, this study aimed to compare single- versus multiple-stoma endoscopic fenestration for treating Galassi type III MCFACs. METHODS: This retrospective study included 86 patients with Galassi type III MCFACs treated with endoscopic fenestration. Single-stoma fenestration to the basal cistern was performed in 37 cases, whereas multiple-stoma fenestration to the basal cistern and the carotid cistern was performed in 49 cases. Clinicoradiologic profiles and follow-up data were analyzed. RESULTS: The rate of symptom relief was 83.7% (72/86), and the rate of cyst shrinkage was 96.5% (83/86). Postoperative ipsilateral subdural effusion, which was significant (p = 0.042), and noninfectious fever were the two most common complications in the single- and multiple-stoma groups. No significant differences in intraoperative nerve injury, vascular injury, proportion of cases with cyst reduction, and symptom remission rate were observed between the two groups. The rates of cyst recurrence and secondary surgery in the single-stoma group were higher than those in the multiple-stoma group, although the difference was not significant. CONCLUSION: Endoscopic fenestration is an effective and minimally invasive approach for treating Galassi type III MCFACs. Single- and multiple-stoma endoscopic fenestrations have the same curative effect.

Identification and Verification of SLC27A1, PTBP1 and EIF5A With Significantly Altered Expression in Aggressive Pituitary Adenomas.

Background: Aggressive pituitary adenoma encircling the internal carotid artery has a poor clinical prognosis because of a high surgical risk and a high recurrence rate. This seriously affects patients' quality of life and yet there is no effective medical treatment. The European Diagnostic Guidelines have recommended the use of temozolomide (TMZ) for these aggressive pituitary adenomas, but the treatment remission rate has been less than 50%. Methods: In this study, transcriptome sequencing of pituitary tumour tissues and TMZ-treated pituitary tumour cell lines were employed to explore the significance gene expressions affecting the efficacy of TMZ treatment for pituitary tumours. To clarify the roles of these gene expressions, six adult patients with pituitary adenomas treated in Tiantan Hospital from 2015 to 2020 and a pituitary adenoma cell line (Att20 sensitive to TMZ treatment) were analyzed by mRNA transcriptome sequencing. The differentially expressed genes were assayed by analyzing the sequencing results, and the expression level of these genes was further verified by immunohistochemistry. In addition, Ki67, VEGF, and p53 of the tumour tissues were also verified by immunohistochemistry. Results: In tumour tissues, mRNA sequencing showed that PTBP1 and EIF5A were significantly overexpressed in primary pituitary adenomas and SLC27A1 was significantly overexpressed in aggressive pituitary adenomas. Also in the pituitary adenoma cell line (AtT20), SLC27A1 expression levels were suppressed by TMZ treatment. Subsequent immunohistochemistry confirmed the sequencing results. Conclusion: High expression of SLC27A1 and low expression of EIF5A and PTBP1 may be potential indicators to predict the progression of aggressive pituitary adenomas, and patients with high SLC27A1 subtype may be sensitive to TMZ in clinical treatments.

Suprasellar cysts: clinical presentation, surgical indications, and optimal surgical treatment.

BACKGROUND: To describe the clinical presentation of suprasellar cysts (SSCs) and surgical indications, and compare the treatment methods of endoscopic ventriculocystostomy (VC) and ventriculocystocisternotomy (VCC). METHODS: We retrospectively reviewed the records of 73 consecutive patients with SSC who were treated between June 2002 and September 2009. Twenty-two patients were treated with VC and 51 with VCC. Outcome was assessed by clinical examination and magnetic resonance imaging. RESULTS: The patients were divided into five groups based on age at presentation: age less than 1 year (n = 6), 1-5 years (n = 36), 6-10 years (n = 15), 11-20 years (n = 11), and 21-53 years (n = 5). The main clinical presentations were macrocrania (100%), motor deficits (50%), and gaze disturbance (33.3%) in the age less than 1 year group; macrocrania (75%), motor deficits (63.9%), and gaze disturbance (27.8%) in the 1-5 years group; macrocrania (46.7%), symptoms of raised intracranial pressure (ICP) (40.0%), endocrine dysfunction (40%), and seizures (33.3%) in the 6-10 years group; symptoms of raised ICP (54.5%), endocrine dysfunction (54.5%), and reduced visual field or acuity (36.4%) in the 11-20 years group; and symptoms of raised ICP (80.0%) and reduced visual field or acuity (40.0%) in the 21-53 years group. The overall success rate of endoscopic fenestration was 90.4%. A Kaplan-Meier curve for long-term efficacy of the two treatment modalities showed better results for VCC than for VC (p = 0.008). CONCLUSIONS: Different age groups with SSCs have different main clinical presentations. VCC appears to be more efficacious than VC.

Assessment of endoscopic treatment for middle cranial fossa arachnoid cysts.

BACKGROUND: Endoscopic cystocisternotomy is one of three surgical methods used to treat middle cranial fossa arachnoid cysts. There is debate about which method is the best. OBJECTIVE: The aim of this study is to evaluate the effectiveness and safety of endoscopic cystocisternotomy for treatment of arachnoid cysts of the middle cranial fossa. METHODS: Thirty-two patients with arachnoid cysts of the middle cranial fossa who had undergone endoscopic cystocisternal fenestration between 2004 and 2009 were studied retrospectively. Data were obtained on clinical and neuroradiological presentation, indications to treat, surgical technique, complications, and the results of clinical and neuroradiological follow-up. RESULTS: Among the 27 patients with symptoms before surgery, 8 had disappearance of symptoms and 17 had improvement of symptoms. The cyst was reduced in size or it completely disappeared in 24 (75%) patients. The incidence rate of complications was 18.8%. CONCLUSIONS: Endoscopic cystocisternal fenestration is an effective treatment for symptomatic arachnoid cysts of the middle cranial fossa and should be the initial surgical procedure.

[Associativity between differentiation potential and VEGF secretory capability of immortalized human mesenchymal stem cells].

OBJECTIVE: To investigate the multi-differentiation potential and VEGF secretory volume of monoclonal immortalized human mesenchymal stem cells (hMSC-TERT) and to determine the relationship between them. METHODS: Monoclonal hMSC-TERT were isolated using limiting dilution. The growth curves of them were detected by method of MTT. ELISA was used to detect the concentration of VEGF in the supernatant of those monoclonal hMSC-TERT. Their adipocytic, osteogenic, neuronal differentiation potential in vitro were determined by Oil Red O staining, Von Kossa staining and immunocytochemistry for Tubulin-ßIII antibody. Those Monoclonal hMSC-TERT were transplanted into the subcutaneously of severe combined immunodeficiency (SCID) mice. The grafts of those cells were removed and analyzed by immunohistochemistry for pathologic tissue markers to discover the multi-differentiation potential of those monoclonal hMSC-TERT in vivo. RESULTS: There was statistically significant difference between different monoclonal hMSC-TERT in mult differentiation potential and the decreased concentration of VEGF in the supernatant of those monoclonal hMSC-TERT from the 3(rd) day to the 5(th) day. The positive rates of CK in grafts formed by those monoclonal hMSC-TERT in SCID mice were direct correlation with the decreased concentration of VEGF in the supernatant of those cells. CONCLUSION: The secretory capability of VEGF of those monoclonal hMSC-TERT may direct correlation with the epithelial differentiation potential of those cells.

[Application of neuroendoscope in the treatment of skull base chordoma].

OBJECTIVE: To further explore the application, approach, indication and prognosis of neuroendoscope treatment for skull base chordoma. METHODS: A total of 101 patients of skull base chordoma were admitted at our hospital from May 2000 to April 2010. There were 59 males and 42 females. Their major clinical manifestations included headache, cranial nerve damage and dyspnea. They were classified according to the patterns of tumor growth: Type I (n = 13): tumor location at a single component of skull base, i. e. clivus or sphenoid sinus with intact cranial dura; Type II (n = 56): tumor involving more than two components of skull e. g clivus, sphenoid and nasal/oral cavity, etc. But there was no intracranial invasion; Type III (n = 32) : tumor extending widely and intradurally forming compression of brain stems and multiple cranial nerves. Based on the types of chordoma, different endoscopic approaches were employed, viz. transnasal, transoral, trans-subtemporal fossa and plus microsurgical craniotomy for staging in some complex cases. RESULTS: Among all patients, total resection was achieved (n = 19), subtotal (n = 58) and partial (n = 24). In partial resection cases, 16 cases were considered to be subtotal due to a second-stage operation. Most cases had conspicuous clinical improvements. Self-care recovery within one week post-operation accounted for 58.4%, two weeks 30.7%, one month 6.9% and more than one month 1.9%. Postoperative complications occurred in 13 cases (12.8%) and included CSF leakage (n = 4) cranial nerve palsy (n = 5), hemorrhagic nasal wounds (n = 3) and delayed intracranial hemorrhage (n = 1). All of these were cured or improved after an appropriate treatment. A follow-up of 6 - 60 months was conducted in 56 cases. CONCLUSION: Early detection and early treatment are crucial for achieving a better outcome in chordoma. Neuroendoscopic treatment plays an important role in managing those complicated cases. Precise endoscopic techniques plus different surgical approaches and staging procedures are required to improve the post-operative quality of life for patients.

LncRNA MEG8 promotes TNF-α expression by sponging miR-454-3p in bone-invasive pituitary adenomas.

There are few studies on the mechanism of pituitary adenoma (PA) destroying bone. The current study aimed to investigate the role of MEG8/miR-454-3p/TNF-α in bone-invasive pituitary adenomas (BIPAs). In this study, we report that lncRNA MEG8 and TNF-α are upregulated in BIPA tissues while miR-454-3p is downregulated, which is associated with poor progression-free survival (PFS). Functional assays revealed the role of up-regulated MEG8 and down-regulated miR-454-3p in promoting bone destruction. Mechanistically, MEG8 promotes TNF-α expression by sponging miR-454-3p, which ultimately leads to the occurrence of bone destruction. The mechanism is confirmed in vivo and in vitro. Therefore, our data illustrated a new regulatory mechanism of MEG8/miR-454-3p/TNF-α in BIPAs. It may provide a useful strategy for diagnosis and treatment for BIPA patients.

Clinical features, radiological profiles, pathological features and surgical outcomes of pituicytomas: a report of 11 cases and a pooled analysis of individual patient data.

BACKGROUND: Pituicytoma is an extremely rare low-grade glial tumor that is closely related to the neurohypophysis axis. Most studies of pituicytomas include only several cases. To better understand this disease, we reviewed a series of cases of pituicytomas. The diagnosis and treatment of pituicytoma must be further elucidated. METHODS: Eleven patients with pituicytoma admitted to Beijing Tiantan Hospital from 2012 to 2019 were selected. The clinical features, including radiological and histological examination, surgical records and prognosis were reviewed. Sixty-eight other previously published cases of pituicytoma also were used to analyze the predictive factors for the results. The Cox regression model was used for univariate and multivariate analyses. RESULTS: Our patients included 5 males (45.5%) and 6 females (54.5%), with a mean age of 49.3 years. The tumor was located in the suprasellar region in 5 patients (45.5%), intrasellar region in 4 patients (36.4%), and intrasellar-suprasellar region in 2 patients (18.2%). All patients were misdiagnosed with other common tumors in the sellar region before the operation. During the operation, gross total resection (GTR) of the tumor was achieved in 6 patients (54.5%), and subtotal resection (STR) was achieved in 5 patients (45.5%). The mean progression-free survival (PFS) time was 29.82 months. Tumor progression after surgical resection occurred in 4 patients (36.4%). Among them, 60.0% of the patients (cases 4, 5, 7) with STR experienced progression, while 16.7% of the patients (case 2) with GTR experienced progression. Combined with the 68 cases in the literature, GTR was an independent risk factor for PFS time (P < 0.05). CONCLUSIONS: Pituicytomas are more common in middle-aged people and the sellar region. The clinical manifestations of pituicytomas are different, but no diagnostic clinical features have been identified other than an abnormally abundant blood supply. Currently, GTR is the best approach for the treatment of pituicytomas. More patients and longer follow-up periods were needed to further elucidate the biological features of pituicytomas.

[Microsurgical treatment of giant cell tumor of skull].

OBJECTIVE: To explore the diagnosis and microsurgical treatment of giant cell tumor (GCT) of skull. METHODS: The investigators reviewed the clinical features, operative approach and prognosis of 23 consecutive cases of GCT of skull operated at our department between July 2000 and November 2008. RESULTS: Headache was the most common presenting symptom (86.96%) found among the patients with GCT of skull. Besides, the symptoms induced by the consequent lesions of II-XII intracranial nerves were also commonly observed. The mean duration of symptoms was 29.3 months and the mean preoperative KPS (Karnofsky performance scale) was 76 +/- 6. Among these cases, 10 tumors occurred in sphenoid bone, 9 in temporal bone, 3 in posterior fossa and 1 in frontal bone. Gross total resections were achieved in 6 cases, subtotal resections in 10 cases and partial resections in 7 cases. There was no operative death case. The follow-up data of 18 patients (78.26%) were collected with a mean follow-up duration of 35.5 months. All patients lived in normal postoperative life. CONCLUSION: GCT of skull is a generally low-degree malignancy showing a local bone invasion mostly in sphenoid and temporal bones.

[Using MR PC cine to evaluate CSF dynamics in neuroendoscopic third ventriculostomy:an analysis of 146 cases].

OBJECTIVE: To evaluate the change of CSF dynamics using MR PC Cine for neuroendoscopic third ventriculostomy. METHODS: 146 cases of hydrocephalus were treated by neuroendoscopic third ventriculostomy including 36 cases checked with MR PC Cine study randomly. The successful result was assessed by clinical symptom and imaging study. All the patients were given 3 months to 1 year follow-up. RESULTS: The symptoms of 121 (83%) patients were recovered soon. CT, MRI and MR PC Cine demonstrated the CSF velocity, flow rate and dynamics change to the normal level compared with preoperative check. The effective rate of this group was 75.3% with one year follow-up. CONCLUSION: The method of MR PC Cine to evaluate the CSF dynamics result for neuroendoscopic third ventriculostomy is simple, fast and safe. It is worth the clinical application.

[Endoscopic treatment of middle fossa arachnoid cysts].

OBJECTIVE: To investigate and evaluate the effectiveness of neuroendoscopic therapy for arachnoid cysts of middle cranial fossa. METHODS: From January 2004 to June 2009, 32 patients with arachnoid cysts of middle cranial fossa who were treated with endoscopic cystocisternal fenestration were retrospectively analyzed. There were 21 male patients and 11 female patients, aged from 6 months to 39 years. The clinical and neuroradiological presentation, indications, surgical technique, complications, and clinical and neuroradiological follow-up were analyzed. RESULTS: The cysts were reduced in size in 20 patients and completely disappeared in 4 patients. For the 27 patients with symptoms before operation, the symptoms disappeared in 8 cases and improved in 17 cases after operation. There were asymptomatic subdural hydroma in 4 patients, intracranial infection and incision cerebro-spinal fluid leakage in 1 patient respectively. The complication incidence rate was 18.8%. CONCLUSIONS: Endoscopic fenestration is an effective treatment for symptomatic arachnoid cysts of middle cranial fossa and could be performed as the first surgical choice for these patients.

Correlation of alternative splicing of the D2 dopamine receptor mRNA and estrogen receptor mRNA in the prolactinomas and gonadotrope tumors.

OBJECTIVE: Estradiol (E2) acts to modulate the ratio of two dopamine D2 receptor isoforms (D2L/D2S) by the nuclear estrogen receptor (ER) and to reduce dopamine's inhibitory action on PRL secretion. Here we demonstrate the correlation between the expression of ER mRNA and D2R mRNA isoforms in pituitary neoplasms cells. METHODS: Twenty-four human pituitary adenomas (14 prolactinomas and 10 gonadotrope tumors) were examined for the expression of both ER mRNA and D2R mRNA by means of semi-quantitative RT-PCR analysis. RESULTS: No significant difference was found in ERbeta mRNA expression levels between prolactinomas and gonadotrope tumors (P = 0.871), but there was a significant difference in the expression of ERalpha mRNA (P = 0.003). The significant difference was found between the two pituitary adenomas types in both levels of D2S and D2L mRNA expression (P = 0.036 and 0.007 respectively). Furthermore, both levels of expression in prolactinomas were significantly higher than that in gonadotrope tumors. Additionally, a negative correlation between D2S and ERalpha mRNA expression and a positive correlation between D2L and ERalpha mRNA expression were found in these tumors. CONCLUSION: This study for the first time shows a good correlation between expression of ER and D2R isoforms in prolactinomas and gonadotrope tumors. Reducing the amount of the ERalpha in neoplasm cells can alter the ratio of D2L/D2S, which may increase the drug sensitivity of pituitary adenomas.

Effect of bone marrow stromal cell-conditioned medium on the glutamate uptake of peroxide-injured astrocytes.

We aimed to investigate the effect of bone marrow stromal cell-conditioned medium (BCM) on glutamate uptake of peroxide (H(2)O(2))-injured astrocytes. Bone marrow stromal cells (BMSC) were isolated from rat bone marrow. Confluent BMSC cultures were incubated with serum-free Dulbecco's Modified Eagle's Medium to create the BCM. Astrocytes were isolated from 1-day-old rats. H(2)O(2)-injured astrocytes were cultured in BCM (experimental group) or serum-free medium (control group). The labeled glutamate ((3)H-L-glutamate) uptake by H(2)O(2)-injured astrocytes with or without BCM was compared after 1 and 3 days. We found that astrocytes cultured in BCM exhibited increased glutamate uptake compared to those cultured in serum-free medium following H(2)O(2)-induced injury (p<0.01) and concluded that BCM increased the glutamate uptake capability of H(2)O(2)-injured rat astrocytes. The therapeutic benefits associated with BMSC transplantation following brain injury might be partly due to increased glutamate uptake by astrocytes.

[Effect of monoclonal or different cell seeding densities on the differentiation potential of immortalized human mesenchymal stem cells in vitro and in vivo].

OBJECTIVE: To determine whether monoclonality or different cell seeding densities could influence the differentiation potential of immortalized human mesenchymal stem cells (hMSC-TERT) and to find an effective cultural method of hMSC-TERT in vitro. METHODS: From the parental hMSC-TERT cell line, we derived 30 monoclonal cell lines and two independent cell lines based on different plating densities during expansion in culture. Their adipocytic, osteogenic, neuronal differentiation potential in vitro and multidirectional differentiation potential in vivo were analyzed by immunohistochemistry for pathologic tissue markers. RESULTS: Monoclonal cell lines and the cell line derived at low seeding density had a lower differentiation potential in vitro than the cell line derived at higher cell seeding density. The differentiation potential of monoclonal hMSC-TERT cells were dissimilar. Some of monoclonal hMSC-TERT lines expressed epithelial differentiation potential in vivo while the parental hMSC-TERT cells line did not. CONCLUSION: Multiclonal hMSC-TERT cells cultured in high seeding density can keep the differentiation potential, cloning the hMSC-TERT cells before transplantation to find the special clones for special purpose of transplantation.